A variant form of laminin is responsible for the neurite outgrowth-promoting activity in conditioned medium from a squamous carcinoma cell line

Abstract

We examined the effects of conditioned medium (CM) obtained from a squamous carcinoma cell line, termed SAS, on chick sympathetic neuritic outgrowth. Neurons grown on a substratum coated with CM extended their neurites. Antisera raised against human laminin and mouse EHS laminin immunoprecipitated the neurite outgrowth-promoting factor in CM. The most purified fraction of CM contained a 740-kDa protein which reacted with the anti-laminin sera and was composed of three polypeptides of 330 kDa, 215 kDa and 195 kDa. The 215-kDa and 195-kDa polypeptides, but not the 330-kDa polypeptide were shown to be antigenically related to mouse laminin and human laminin by immunoblotting. The presence of merosin M chain in SAS cells was ruled out by using the method of reverse transcription polymerase chain reaction (RT-PCR). These results suggest that the 740-kDa protein is a laminin variant having a novel heavy chain of 330 kDa and is responsible for neurite outgrowth-promoting activity in CM.